• Title/Summary/Keyword: Enzyme Efficiency

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Effect of Zona Hardening on In Vitro Fertilization in Mouse Oocytes I. Analysis of Zona Soluble Properties with Proteolytic Enzymes (생쥐난자에 있어서 투명대 경화 현상이 체외수정에 미치는 영향 I. 단백분해효소에 의한 투명대의 용해성 분석)

  • 이상진;이상민;지희준;장경환;정형민;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.93-101
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    • 1993
  • These experiments were carried out to investigate whether zona hardening affect the efficiency of in vitro fertilization in mouse oocytes. The soluble properties for zona pellucida of oocytes matured in vivo, aged oocytes, and ovarian oocytes matured in vitro have been analyzed with proteolytic enzyme, 3mg/ml of $\alpha$-chymotrypsin. The mean solubility(t50) for the zona of unfertilized oocytes, oocytes not fertilized at the first inseminati and in vitro produced zygotes were 10.1, 20.3 and 32.3min., respectively. The t50 for zona lysis of fertilized oocytes was significantly difference than those observed for unfertilized oocytes and oocytes not fertilized at the first insemination(P<0.01). In addition, the t50 of zona in ovulated oocytes with and without cumulus cells incubated for 0, 3, 6, 9 and 12 hr in vitro, t50 were 13.9, 11.1, 20.7 and 28.0min., and 22.3, 21.0, 30.0 and 33.5min., respectively. In these experiments, the zona pellucida showed a gradual increase in resistance to dissolution by $\alpha$-chyjotrypsin with in vitro aging for more than 6 hrs. This effect was greater in cumulus-free as compared to cumulus-intact oocytes. Finally, in cumulus-intact and cumulus-free ovarian oocytes matured for 0, 5, 10 and 15 hr in vitro the t50 of zona pellucida were 3.0, 10.6, 18.4 and 24.5 min., and 3.0, 14.0, 26.2 and 32.0 min., respectively. Clear differences in solubility between the zona pellucida of oocytes matured in vivo and in vitro. This data were found suggest that under in vitro conditions there is a gradual change in the soluble properties of the zona pellucida, particularly in the absence of the cumulus cells.

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The Effect of Glucose and Glucose Transporter on Regulation of Lactation in Dairy Cow

  • Heo, Young-Tae;Park, Joung-Jun;Song, Hyuk
    • Reproductive and Developmental Biology
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    • v.39 no.4
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    • pp.97-104
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    • 2015
  • Glucose is universal and essential fuel of energy metabolism and in the synthesis pathways of all mammalian cells. Glucose is the one of the major precursors of lactose synthesis using glycolysis result in producing milk fat and protein. During the milk fat synthesis, lipoprotein lipase (LPL) and CD36 are required for glucose uptake. Various morecules such as acyl-CoA synthetase 1 (ACSL1) activity of acetyl-CoA synthetase 2 (ACSS2), ACACA, FASN AGPAT6, GPAM, LPIN1 are closely related with milk fat synthesis. Additionally, glucose plays a major role for synthesizing lactose. Activations of lactose synthesize enzymes such as membranebound enzyme, beta-1,4-galactosyl transferase (B4GALT), glucose-6-phosphate dehydrogenase (G6PD) are changed by concentration of glucose in blood resulting change of amount of lactose production. Glucose transporters are a wide group of membrane proteins that facilitate the transport of glucose over a plasma membrane. There are 2 types of glucose transporters which consisted facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Among them, GLUT1, GLUT8, GLUT12, SGLT1, SGLT2 are main glucose transporters which involved in mammary gland development and milk synthesis. However, more studies are required for revealing clear mechanism and function of other unknown genes and transporters. Therefore, understanding of the mechanisms of glucose usage and its regulation in mammary gland is very essential for enhancing the glucose utilization in the mammary gland and improving dairy productivity and efficiency.

Expression of Antioxidant Isoenzyme Genes in Rice under Salt Stress and Effects of Jasmonic Acid and ${\gamma}$-Radiation

  • Kim, Jin-Hong;Chung, Byung-Yeoup;Baek, Myung-Hwa;Wi, Seung-Gon;Yang, Dae-Hwa;Lee, Myung-Chul;Kim, Jae-Sung
    • Journal of Applied Biological Chemistry
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    • v.48 no.1
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    • pp.1-6
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    • 2005
  • Analysis of chlorophyll (Chl) fluorescence implicated treatment of 40 mM NaCl decreased maximal photochemical efficiency of photosystem II (PSII) (Fv/Fm), actual quantum yield of PSII (${\Phi}_{PSII}$), and photochemical quenching (qP) in rice, but increased non-photochemical quenching (NPQ). Decreases in Fv/Fm, ${\Phi}_{PSII}$, and qP were significantly alleviated by $30\;{\mu}M$ jasmonic acid (JA), while NPQ increase was enhanced. Transcription levels of antioxidant isoenzyme genes were differentially modulated by NaCl treatment. Expression of cCuZn-SOD2 gene increased, while those of cAPXb, CATb, and CATc genes decreased. JA prevented salt-induced decrease of pCuZn-SOD gene expression, but caused greater decrease in mRNA levels of cAPXa and Chl_tAPX genes. Investigation of vacuolar $Na^+/H^+$ exchanger (NHX2) and 1-pyrroline-5-carboxylate synthetase (P5CS) gene expressions revealed transcription level of NHX2 gene was increased by JA, regardless of NaCl presence, while that of P5CS gene slightly increased only in co-presence of JA and NaCl. Unlike JA, ${\gamma}$-radiation rarely affected expressions of antioxidant isoenzyme, NHX2, and P5CS genes, except for increase in mRNA level of Chl_tAPX and decrease in that of pCuZn-SOD. These results demonstrate enhanced salt-tolerance in JA-treated rice seedlings may be partly due to high transcription levels of pCuZn-SOD, NHX2, and P5CS genes under salt stress.

Effects of Biochar on Soil Quality and Heavy Metal Availability in a Military Shooting Range Soil in Korea

  • Lee, Sung-Eun;Ahmad, Mahtab;Usman, Adel A.R.A.;Awad, Yasser M.;Min, Sun-Hong;Yang, Jae-E;Lee, Sang-Soo;Ok, Yong-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.67-77
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    • 2011
  • Heavy metal remediation in shooting range soil is a challenge over the world. The excessive Pb accumulation in the soil can deteriorate soil quality and fertility. The objectives of this research were to evaluate the efficiency of biochar (BC) in improving the physicochemical and biological properties of the soil and to evaluate its effect on Pb availability in a military shooting range soil. Sandy loam soil was collected from shooting range of Gyeonggi Province, South Korea and was incubated for 30 days with different application rates (0-30% w $w^{-1}$) of BC. The results showed that the addition of BC increased aggregate stability, nitrogen (N) and phosphorus (P) contents, and enzyme activities in soil. Sequential extraction showed that residual and organic bound fractions in the soil amended with BC increased by 33.1 and 16.7%, respectively, and the exchangeable fraction decreased by 93.7% in the soil amended with BC, compared to the unamended soil. We concluded that the application of BC could not only improve physicochemical and biological soil qualities but also stabilize Pb in a shooting range soil.

Protein Quality Evaluation of Cooked Monkfish (Lophiomus setigerus) Meats

  • Jeung Young-Ae;Ryu Hong-Soo;Shin Eun-Soo;Mun Sook-Im
    • Fisheries and Aquatic Sciences
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    • v.6 no.4
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    • pp.165-171
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    • 2003
  • To investigate the effect of cooking methods on protein quality of domestic fresh monkfish meat (FMM) and imported frozen monkfish meat (IMM), in vitro protein qualities were determined by amino acid anlysis, trypsin indigestible substrate (TIS) formation, and protein digestibility using the four-enzyme method. Crude protein contents of the boiled FMM and IMM were $90\%$ of the dry base, which were higher than fresh FMM $(82\%)$ and IMM $(84\%)$. Profiles of total amino acid in FMM and IMM were not changed by cooking methods. Total free amino acid contents decreased to $ 29.0-33.6\%$ for boiled $(l00^{\circ}C,\;10 min)\;and\;24\%$ for steamed $(100^{\circ}C,\;10\;min)$ samples. In vitro protein digestibilities of boiled and steamed FMM incnased $86.6-86.8\%$, compared to raw IMM $(82.9\%)$, boiled and steamed IMM $85.1-85.5\%$ and raw IMM $(83.6\%)$. TIS of FMM (23.6 mg/g solid) and IMM (15.9 mg/g solid) showed no significant (p<0.05) difference in cooking methods. The C-PERs (computed protein efficiency ratio) of boiled FMM (2.63) and IMM (2.50) were significantly higher (<0.05) than raw (1.97) and steamed FMM(1.97) and IMM(1.94). These results demonstrate that boiling of FMM and IMM improves protein digestibility and C-PER when compared to steamed FMM and IMM. Therefore, boiling could be an excellent means to maintain high-protein quality of monkfish meat. Also, the cooking method may be applicable to the preparation of monkfish stew without any loss of free amino acids.

Characterization of the Folding Structure of 3'-end of Lactate Dehydrogenase A-mRNA Isolated from Hormone Stimulated Rat $C_{6}$ Glioma cell culture (홀몬으로 처리된 쥐의 $C_{6}$ glioma 세포배양으로부터 분리된 낙산탈수소 효소 A-mRNA의 3'-말단의 2차 구조)

  • 배석철;이승기
    • Korean Journal of Microbiology
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    • v.25 no.2
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    • pp.94-102
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    • 1987
  • Rat liver LDH A-cDNA has been isolated from a .lambda.gt11-rat lover cDNA library and partially characterized. The size of the isolated rat liver LDH A-cDNA if shown to be 1.6Kb and restriction enzyme sites for the rat liver LDH A-cDNA are also mapped. 682-nucleotide sequence coding for 3'-end of rat liver LDH A-cDNA has been analyzed and compared to the nucleotide sequence of the same region of rat $C_6$-glioma cell LDH A-cDNA which has been cloned from the hormonally stimulated cell cultures. The result shows that 177 nucleotide sequences coding for the C-terminal 59-amino acids are identical but 505 nucleotide sequences of 3'-nontranslated region of the two LSH A-cDNA exhibit characteristic differences in thier nucleotide sequences. Computer analysis for the folding structures for 3'-end 400 nucleotide sequences of the two LDH A-cDNA shows a possibility implying that the two LDH A-mRNAs isolated from different tissues of rats may have different half life and therefore their translational efficiency may be different. It has been previously demonstrated that isoproterenol stimulated rat $C_6$ -glioma cell cultures produce LDH A-mRNA showing 2 to 3-fold longer half life in comparison to that of noninduced LHD A-mRNA. The result therefore support for the idea that hormonally stimulated rat $C_6$-glioma cells may produce LDH A-mRNA containing different nucleotide sequences at the 3'-end nontranslated region by which the hormonally induced LDH A-mRNA could have more stable secondary mRAN structure in comparison to that of noninduced LDH A-mRNA.

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Analysis of Improved Shear Stiffness and Strength for Sandy Soils Treated by EICP (EICP 방법으로 처리된 사질토의 전단 강성도 및 강도 증가 분석)

  • Song, Jun Young;Ha, Seong Jun;Jang, Jae Won;Yun, Tae Sup
    • Journal of the Korean Geotechnical Society
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    • v.36 no.1
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    • pp.17-28
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    • 2020
  • This study presents the experimental results of ground improvement efficiency induced by enzyme-induced carbonate precipitation (EICP) in soils. First, the optimal mixture ratio of EICP solution was determined by comparing the amount of induced carbonate depending on the different ratio among urea, CaCl2, and urease. Next, we evaluated the shear stiffness and strength of EICP-treated sandy soil by performing shear wave velocity measurement and triaxial shear test. Furthermore, induced carbonate in treated soil was visually investigated by X-ray CT and SEM analysis. The results showed that the maximum shear stiffness evolved 19~30 times after 6 hours of reaction time compared with non-treated sands. Also, the cohesion and the friction angle tended to increase and decrease, respectively, as the amount of induced carbonate increased.

The Effects of Phenolic Radical Carriers on the Enzymatic. Bleaching of Kraft Pulp (Kraft펄프의 효소표백반응에 미치는 페놀라디칼 전달체의 영향)

  • 류근갑
    • KSBB Journal
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    • v.10 no.2
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    • pp.183-190
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    • 1995
  • The effects of enzymatic pretreatments on the bleaching of kraft pulp were studied. The kappa number of pulp samples which represents the lignin content of pulp decreased by 25.2% by the pretreatments with xylanase(EC 3.2.1.8, Pulpzyme HB) while it decreased by 13.7% without enzyme pretreatments after the extraction of the pretreated pulp samples in 1N NaOH. To enhance the effects of enzymatic pretreatment on the bleaching of kraft pulp, phenols were used as radical carriers with the simultaneous use of peroxidase(EC 1.11.1.7, Novozyme 502), $H_2O_2$, and xylanase. Guaiacol (1mM) was most effective by decreasing the kappa number by 29.6% when a low initial concentration of $H_2O_2$ (0.1mM) was used. The use of either a higher initial concentration of $H_2O_2$ or phenols lacking electron donating substituents such as phenol and p-chloyophenol, however, decreased the efficiency of enzymatic pretreatment indicating that the production rate and the stability of phenolic radicals are important parameters.

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Oxidative Stress and Antioxidant Activities of Intertidal Macroalgae in Korea

  • Park, Jung-Jin;Han, Tae-Jun;Choi, Eun-Mi
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.313-320
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    • 2011
  • The oxidative stress level and antioxidant activities in two green algae (Ulva pertusa and Ulva linza), two brown algae (Agarum cribrosum and Dictyota dichotoma), and three red algae (Grateloupia lanceolata, Carpopeltis affinis, and Gracilaria verrucosa) collected from intertidal regions of Korea were assessed. In the two green algae, although the total glutathione content was not as high as that of the brown algae, the glutathione pool was extremely reduced, and the glutathione reductase (GRd)/glutathione peroxidase (GPx) activity ratio was high, which apparently plays an important role for protection against oxidative damage, as manifested by low lipid peroxidation. In the brown algae, which exhibited a low lipid peroxidation level that was comparable to the green algal species, the highest glutathione content, together with high GPx activity, appears to be the most important factor in their antioxidant protection. The red algal species exhibited extremely high lipid peroxidation levels. They also contained the lowest and most oxidized glutathione among the species, as well as the lowest GRd activity. In spite of the marked difference in the glutathione content, the significant difference in the activity of ${\gamma}$-glutamylcysteine ligase, the rate limiting enzyme for glutathione synthesis, among the species was not exhibited. Our results suggest that there is a significant difference in the levels of oxidative stress and antioxidant capacity among the algal species, and that the glutathione system, especially the efficiency of glutathione recycling, plays a vital role in antioxidative protection in algal species.

Preferential Peroxidase Activity of Prostaglandin Endoperoxide H Synthase for Lipid Peroxides

  • Yun, Seol-Ryung;Han, Su-Kyong;Song, In-Seok
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.94-94
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    • 2001
  • Prostaglandin endoperoxide H synthase (PGHS) catalyzes the committed step in prostaglandins and thromboxane A$_2$-- oxygenation of arachidonic acid to the hydroperoxy endoperoxide PGG$_2$, followed by reduction PGG$_2$to the alcohol PGH$_2$. The two reactions by PGHS -- cyclooxygenase and peroxidase -- occur at distinct but structurally and functionally interconnected sites. The peroxidase reaction occurs at a heme-containing active site located near the protein surface. The cyclooxygenase reaction occurs in a hydrophobic channel in the core of the enzyme. Initially a peroxide reacts with the heme group, yielding Compound I and an alcohol derived from the oxidizing peroxide. Compound I next undergoes an intramolecular reduction by a single electron traveling from Tyr385 along the peptide chain to the proximal heme ligand, His388, and finally to the heme group. Following the binding of arachidonic acid, Tyr385 tyrosyl radical initiates the cyclooxygenase reaction by abstracting the 13-pro(5) hydrogen atom to give an arachidonyl radical, which sequentially reacts with two molecules of oxygen to yield PGG$_2$. In order to characterize PGHS peroxidase active site, we examined various lipid peroxides with purified recombinant ovine PGHS proteins and determined the rate constants. The results have shown that twenty-carbon unsaturated fatty acid hydroperoxides have similar efficiency in peroxidation by PGHS, irrespective of either the location of hydroperoxy group or the number of double bonds. It was also confirmed by the subsequent study with PGHS peroxidase active site mutants.

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